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Efficient isolation of high-quality RNA from lotus Nelumbo nucifera ssp nucifera tissues.
Zhang, Y J; Hao, X Y; Liang, Z S; Ke, W D; Guo, H B.
Afiliação
  • Zhang YJ; Shaanxi Key Laboratory of Molecular Biology in Agriculture, College of Life Sciences, Northwest A & F University, Yangling, PR China.
Genet Mol Res ; 12(1): 223-9, 2013 Jan 24.
Article em En | MEDLINE | ID: mdl-23408408
Nelumbo nucifera is widely used as food, as an ornamental, in medicine, and as packing material; it is also reported to have anti-HIV effects and antioxidant capacity. We sought an improved method for extracting high-quality total RNA from different tissues of N. nucifera. Four methods for RNA extraction were assessed for their ability to recover high-quality RNA applicable for evaluation of polyphenol oxidase (PPO) gene expression profiles. The recovery and quality of the RNA obtained from five different tissues by the best CTAB-LiCl method were evaluated through UV light absorbance. Both A(260)/A(280) and A(260)/A(230) absorbance ratios were more than 2.0; the yield ranged from 59.87 to 163.75 µg/g fresh weight. The brightness of the 28S band was approximately twice that of 18S; the latter was also considered as high-quality RNA. The PPO gene fragment (606 bp) was successfully amplified by RT-PCR, demonstrating the integrity of the isolated RNA. The relative expression levels of the PPO gene based on RT-PCR in five tissues of lotus were: rhizome buds (2.66), young leaves (2.42), fresh cut rhizome (2.02), petals (1.80), and petiole (1.65), using housekeeping gene ß-actin as an internal control. We concluded that the total RNA isolated by this protocol is of sufficient quality for molecular applications.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Extratos Vegetais / RNA de Plantas / Nelumbo Idioma: En Revista: Genet Mol Res Assunto da revista: BIOLOGIA MOLECULAR / GENETICA Ano de publicação: 2013 Tipo de documento: Article País de publicação: Brasil

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Extratos Vegetais / RNA de Plantas / Nelumbo Idioma: En Revista: Genet Mol Res Assunto da revista: BIOLOGIA MOLECULAR / GENETICA Ano de publicação: 2013 Tipo de documento: Article País de publicação: Brasil